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使用正丁醇-甲醇-水体系的离心分配色谱法分离木寡糖。

Separation of xylose oligomers using centrifugal partition chromatography with a butanol-methanol-water system.

机构信息

Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, AR 72701, USA.

出版信息

J Ind Microbiol Biotechnol. 2013 Jan;40(1):51-62. doi: 10.1007/s10295-012-1209-7. Epub 2012 Nov 1.

Abstract

Xylose oligomers are the intermediate products of xylan depolymerization into xylose monomers. An understanding of xylan depolymerization kinetics is important to improve the conversion of xylan into monomeric xylose and to minimize the formation of inhibitory products, thereby reducing ethanol production costs. The study of xylan depolymerization requires copious amount of xylose oligomers, which are expensive if acquired commercially. Our approach consisted of producing in-house oligomer material. To this end, birchwood xylan was used as the starting material and hydrolyzed in hot water at 200 °C for 60 min with a 4 % solids loading. The mixture of xylose oligomers was subsequently fractionated by a centrifugal partition chromatography (CPC) with a solvent system of butanol:methanol:water in a 5:1:4 volumetric ratio. Operating in an ascending mode, the butanol-rich upper phase (the mobile phase) eluted xylose oligomers from the water-rich stationary phase at a 4.89 mL/min flow rate for a total fractionation time of 300 min. The elution of xylose oligomers occurred between 110 and 280 min. The yields and purities of xylobiose (DP 2), xylotriose (DP 3), xylotetraose (DP 4), and xylopentaose (DP 5) were 21, 10, 14, and 15 mg/g xylan and 95, 90, 89, and 68 %, respectively. The purities of xylose oligomers from this solvent system were higher than those reported previously using tetrahydrofuran:dimethyl sulfoxide:water in a 6:1:3 volumetric ratio. Moreover, the butanol-based solvent system improved overall procedures by facilitating the evaporation of the solvents from the CPC fractions, rendering the purification process more efficient.

摘要

木二糖低聚物是木聚糖解聚为木糖单体的中间产物。了解木聚糖解聚动力学对于提高木聚糖转化为单体木糖的转化率和最小化抑制性产物的形成从而降低乙醇生产成本非常重要。木聚糖解聚的研究需要大量的木二糖低聚物,如果从商业上获得,这些低聚物非常昂贵。我们的方法包括内部生产低聚物材料。为此,我们使用桦木木聚糖作为起始材料,并在 200°C 的热水中以 4%的固载量水解 60 分钟。随后,通过使用正丁醇:甲醇:水体积比为 5:1:4 的溶剂系统的离心分配色谱(CPC)对木二糖低聚物混合物进行分级。在上升模式下,富含正丁醇的上层(流动相)以 4.89 mL/min 的流速从富含水的固定相洗脱木二糖低聚物,总分级时间为 300 分钟。木二糖低聚物的洗脱时间在 110 到 280 分钟之间。木二糖(DP2)、木三糖(DP3)、木四糖(DP4)和木五糖(DP5)的产率和纯度分别为 21、10、14 和 15mg/g 木聚糖和 95、90、89 和 68%。与之前使用四氢呋喃:二甲基亚砜:水体积比为 6:1:3 的溶剂系统相比,该溶剂系统中木二糖低聚物的纯度更高。此外,基于正丁醇的溶剂系统通过促进 CPC 级分中溶剂的蒸发,使纯化过程更有效率,从而改进了整个过程。

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